Walther F J, Wade A B, Warburton D, Forman H J
Department of Pediatrics, King-Drew Medical Center, UCLA School of Medicine 90059.
Am J Respir Cell Mol Biol. 1991 Apr;4(4):364-8. doi: 10.1165/ajrcmb/4.4.364.
This study tested whether adducts formed by covalent linkage of superoxide dismutase (SOD) or catalase to polyethylene glycol (PEG) could augment SOD and catalase activity in alveolar type II cells and document enhanced resistance to oxidant damage. Alveolar type II cells were isolated from adult, pathogen-free rats. Antioxidant enzymes were added to the medium of cell cultures in various concentrations for periods up to 48 h. Incubation with 500 to 3,000 U of PEG-SOD or 10,000 to 40,000 U of PEG-catalase/10(6) cells produced a dose-response-related increase in intracellular enzyme activity in comparison with controls (untreated or treated with SOD or catalase, inactivated PEG-SOD or PEG-catalase, or PEG alone). Uptake was maximal during the first 4 h. Using fluorescent label (fluorescein isothiocyanate) bound to PEG-catalase, we found intracellular localization of the labeled enzyme. Exposure to H2O2 led to reduced cytotoxicity in cells pretreated with PEG-catalase than in controls. We conclude that supplementation with PEG-SOD or PEG-catalase enhanced the activity of these enzymes in alveolar type II cells and increased their resistance to oxidant stress.
本研究测试了通过超氧化物歧化酶(SOD)或过氧化氢酶与聚乙二醇(PEG)共价连接形成的加合物是否能增强肺泡II型细胞中的SOD和过氧化氢酶活性,并证明对氧化损伤的抗性增强。从成年无病原体大鼠中分离出肺泡II型细胞。将抗氧化酶以不同浓度添加到细胞培养基中,培养长达48小时。与对照组(未处理或用SOD或过氧化氢酶、失活的PEG-SOD或PEG-过氧化氢酶或单独的PEG处理)相比,用500至3000 U的PEG-SOD或10000至40000 U的PEG-过氧化氢酶/10⁶细胞孵育会导致细胞内酶活性呈剂量反应相关增加。摄取在最初4小时内达到最大值。使用与PEG-过氧化氢酶结合的荧光标记(异硫氰酸荧光素),我们发现了标记酶的细胞内定位。与对照组相比,暴露于H₂O₂会导致用PEG-过氧化氢酶预处理的细胞中的细胞毒性降低。我们得出结论,补充PEG-SOD或PEG-过氧化氢酶可增强肺泡II型细胞中这些酶的活性,并增加它们对氧化应激的抗性。